摘要
优化抗肿瘤肽QX3-1融合蛋白在大肠埃希菌中的表达条件,提高表达量。从培养基、诱导温度、诱导剂浓度和诱导时间4个单因素实验入手,优化QX3-1融合蛋白的表达条件,通过聚丙烯酰胺凝胶电泳(SDSPAGE)和Band Scan凝胶分析软件分析QX3-1融合蛋白的表达量。p ED-QX 3-1/BL21重组菌在LB、大豆肉汤、TB和酪蛋白胨(自制) 4种培养基中,于酪蛋白胨培养基中的表达量最高;在20、25和37℃三种诱导温度下,于37℃环境中表达量最高;在1、5、10、15和20 mmol/L乳糖浓度的诱导下,于10 mmol/L乳糖诱导最佳;在30 h的诱导时间内,于第24小时,融合蛋白的表达量最高。从培养基、诱导温度、诱导剂浓度和诱导时间4个变量中筛选出适合QX3-1融合蛋白表达的最佳组合,使融合蛋白的表达量达到了31. 2%,相比未优化前提高了近一倍。
Expression conditions of antitumor peptide QX3-1 fusion protein in E.coli were optimized to increase the expression level. The optimization of QX3-1 fusion protein expression conditions was to begin with four single factors experiment, medium, temperature, concentration of inducer, and induction time, and analyze the expression level of the QX3-1 fusion protein adopting SDS-PAGE and Band Scan gel analysis software. The results showed that the expression level of recombinant strain pED-QX3-1/BL21 on casein medium was the highest among all four media, LB, TSB, TB, and casein （self-made）; the expression level at induction temperature, the one at 37 ℃ was the highest among all three induction temperature, 20 ℃, 25 ℃, and 37 ℃; under the lactose induction, the one induced with 10 mmol/L of lactose was the best among 1 mmol/L, 5 mmol/L, 10 mmol/L, 15 mmol/L, and 20 mmol/L of lactose; and the expression level of fusion protein induced at 24th hours was the highest among those induced within 30 hours. Therefore, the best combination suitable for QX3-1 fusion protein expression was screened among four variables, medium, induction temperature, inducer concentration, and induction time, made the expression level of fusion protein as high as 31.2%, improved nearly one fold as compared with the one that unoptimized.
作者
费迎心
陈豪
李慧婷
吴文惠
王春晓
FEI Ying-xin;CHEN Hao;LI Hui-ting;WU Wen-hui;WANG Chun-xiao(Coll.of Food Sci.＆ Technol.,Shanghai Ocean Uni.,Shanghai 201306)
出处
《微生物学杂志》
CAS
CSCD
2018年第5期78-83,共6页
Journal of Microbiology
基金
国家自然科学基金项目(81502955)
上海市创新行动计划项目(15410722500)
